RNA interference inhibits hepatitis B virus gene expression and replication in HepG2-N10 cells
| Autor: | Song Jie Huang, Jin-Shui Pan, Jing Dong, Tong Cheng, Jian Lin Ren, Ya Pi Lu, Hua Xiu Shi, Ya Mei Lian, Lin Wang |
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| Rok vydání: | 2006 |
| Předmět: |
Gene Expression Regulation
Viral Hepatitis B virus HBsAg Genetic Vectors Biology Transfection Virus Replication medicine.disease_cause Cell Line Small hairpin RNA RNA interference Gene expression medicine Humans Hepatitis B e Antigens RNA Small Interfering Hepatitis B Surface Antigens Gastroenterology RNA Lipids Virology Molecular biology HBeAg Viral replication DNA Viral RNA Viral Indicators and Reagents RNA Interference |
| Zdroj: | Chinese Journal of Digestive Diseases. 7:230-236 |
| ISSN: | 1443-9573 1443-9611 |
| DOI: | 10.1111/j.1443-9573.2006.00268.x |
| Popis: | OBJECTIVE: RNA interference (RNAi) refers to the phenomenon of sequence-specific degradation of homologous mRNA induced by double-stranded RNA. It has been successfully utilized to down-regulate endogenous gene expression or suppress the replication of various pathogens in mammalian cells. In this study, the effect of vector-based small interfering RNA (siRNA) promoted by pSilencer2.0-U6 inhibit hepatitis B virus (HBV) replication in cell culture was evaluated. METHODS: Three fragments of short nucleic acids, respectively, targeting on S, X and C region of HBV genome were inserted into pSilencer vectors after they were annealed with their partly antisense strands. The recombination plasmids were pS, pX and pC. These expression plasmids were transfected into HepG2-N10 cells, a cell line which stably expresses hepatitis B virus surface antigen (HBsAg), hepatitis B virus e antigen (HBeAg) and adw2 subtype Dane particles. The effect of RNAi was evaluated from the changes of DNA, RNA and protein levels. Viral antigens were measured by ELISA. Viral mRNA was analyzed by RT-PCR. The covalent closed circular DNA and genome DNA of HBV secreted into the culture media were measured by quantitative real-time PCR. Analysis of variance was performed for the results. RESULTS: Vector-based RNA interference could potently reduce HBsAg (pS vs pN: 47%, pX vs pN: 30%, and pC vs pN: 25%, P |
| Databáze: | OpenAIRE |
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